Insecticidal proteins

Abstract

Compositions and methods for controlling insect pests are disclosed. In particular, novel insecticidal proteins having toxicity to at least coleopteran insect pests are provided. Nucleic acid molecules encoding the novel insecticidal proteins are also provided. Methods of making the insecticidal proteins and methods of using the insecticidal proteins and nucleic acids encoding the insecticidal proteins of the invention, for example in transgenic plants to confer protection from insect damage, are also disclosed.

1 . A nucleic acid molecule comprising a nucleotide sequence that encodes a protein that is toxic to an insect pest, wherein the nucleotide sequence (a) encodes a protein comprising an amino acid sequence that has at least 80% to at least 99% sequence identity with any of SEQ ID NOs: 1-17, or a toxin fragment thereof; (b) has at least 80% to at least 99% sequence identity with any of SEQ ID NOs:18-40, or a toxin-encoding fragment thereof;
 or (c) is a synthetic sequence of (a) or (b) that has codons optimized for expression in a transgenic organism.   
     
     
         2 . The nucleic acid molecule of  claim 1 , wherein the insecticidal protein comprises an amino acid sequence of any of SEQ ID NOs:1-17, or a toxic fragment thereof. 
     
     
         3 .- 4 . (canceled) 
     
     
         5 . A chimeric gene comprising a heterologous promoter operably linked to the nucleic acid molecule of  claim 1 . 
     
     
         6 .- 11 . (canceled) 
     
     
         12 . A protein that is toxic to an insect pest, wherein the protein comprises (a) an amino acid sequence that has at least 80% to at least 99% sequence identity with an amino acid sequence of any of SEQ ID NOs:1-17, or a toxin fragment thereof; (b) an amino acid sequence that comprises any of SEQ ID NOs:1-17, or a toxin fragment thereof; (c) an amino acid sequence that is encoded by a nucleotide sequence that has at least 80% to at least 99% sequence identity with a nucleotide sequence of any of SEQ ID NOs:18-40, or a toxin-encoding fragment thereof; or (d) an amino acid sequence that is encoded by a nucleotide sequence comprising any of SEQ ID NOs:18-40, or a toxin-encoding fragment thereof. 
     
     
         13 .- 15 . (canceled) 
     
     
         16 . A recombinant vector comprising the chimeric gene of  claim 5 . 
     
     
         17 . A host cell comprising the recombinant vector of  claim 16 , wherein the host cell is a bacterial cell or plant cell. 
     
     
         18 .- 19 . (canceled) 
     
     
         20 . The transgenic plant cell of  claim 17 , wherein the plant cell is a dicot plant cell or a monocot plant cell. 
     
     
         21 . The transgenic plant cell of  claim 20 , wherein (a) the dicot plant cell is selected from the group consisting of a soybean cell, sunflower cell, tomato cell, cole crop cell, cotton cell, sugar beet cell and tobacco cell; or (b) the monocot plant cell is selected from the group consisting of a barley cell, maize cell, oat cell, rice cell, sorghum cell, sugar cane cell and wheat cell. 
     
     
         22 . A transgenic plant or plant part comprising the transgenic plant cell of  claim 21 . 
     
     
         23 . The transgenic pant or plant part of  claim 22  that is a transgenic maize plant or plant part. 
     
     
         24 . An engineered insecticidal protein comprising an amino acid sequence having at least 80% to at least 99% sequence identity to SEQ ID NO:1 and further comprising at least one mutation at a position that corresponds to amino acid positions 1-118 of SEQ ID NO:1 or amino acid positions 33-150 of SEQ ID NO:17. 
     
     
         25 .- 35 . (canceled) 
     
     
         36 . An insecticidal composition comprising the protein of  claim 12  and an agriculturally acceptable carrier. 
     
     
         37 .- 42 . (canceled) 
     
     
         43 . A method for producing an insecticidal protein, comprising culturing the host cell of  claim 17  or an organism comprising the host cell under conditions in which the host cell produces the insecticidal protein. 
     
     
         44 . A method of producing a transgenic plant or plant part having enhanced insect resistance compared to a control plant or plant part, comprising: (a) introducing into a plant or plant part the chimeric gene of  claim 5 , wherein the insecticidal protein is expressed in the plant or plant part, thereby producing a plant or plant part with enhanced insect-resistance. 
     
     
         45 . The method of  claim 44 , wherein the introducing step is achieved by (a) transforming the plant or plant part; or (b) crossing a first plant comprising the chimeric gene with a different second plant. 
     
     
         46 . (canceled) 
     
     
         47 . A method of controlling an insect pest comprising, delivering to the insect pest or an environment thereof an effective amount of the insecticidal protein of  claim 12 . 
     
     
         48 . The method of  claim 47 , wherein the insecticidal protein is delivered through a transgenic plant or by topical application of an insecticidal composition comprising the insecticidal protein. 
     
     
         49 .- 51 . (canceled) 
     
     
         52 . The method of  claim 47 , wherein the insect pest is a coleopteran insect pest. 
     
     
         53 . The method of 52, wherein the coleopteran insect pest is a  Diabrotica  species. 
     
     
         54 . The method of  claim 53 , wherein the  Diabrotica  species is selected from the group consisting of  Diabrotica virgifera virgifera, Diabrotica barberi, Diabrotica undecimpunctata howardi  and  Diabrotica zeae.    
     
     
         55 .- 57 . (canceled)